Search Results (2,552)

Background/Objectives: Global population aging is associated with a rising prevalence of malnutrition among adults aged ≥80 years. Gut dysbiosis is linked to immune decline and impaired nutrient absorption, and aerobic exercise may enhance microbial diversity. This study investigated gut microbiota changes after a 12-week aerobic exercise intervention in octogenarians stratified by nutritional status. Methods: A total of 129 nursing home residents (≥80 years) were classified via the Mini Nutritional Assessment Short-Form (MNA-SF) into a healthy group (HG, MNA-SF ≥ 11) and a malnourished group (MG, MNA-SF < 11). Both groups underwent a 12-week brisk walking intervention (three sessions/week, 1 h/session, 40–60% heart rate reserve). Fecal samples were collected at baseline and post-intervention and were analyzed via shotgun metagenomic sequencing. Results: A total of 36 participants completed the intervention (HG = 17, MG = 19). Within-group baseline-to-post-intervention analysis showed no significant changes in alpha or beta diversity in the MG. However, post-intervention between-group comparison revealed higher microbial richness and diversity in the MG vs. the HG, with enrichment of taxa including Faecalibacterium prausnitzii and Streptococcus salivarius. Functional analysis revealed significant enhancements in metabolic pathways related to amino acid biosynthesis, protein synthesis, and quorum sensing in the MG. In contrast, the HG showed limited shifts in microbial diversity but an increase in species involved in carbohydrate metabolism. Conclusions: After 12 weeks, the malnourished group showed higher post-intervention microbial richness and diversity than the healthy group, with differences in taxonomic and predicted functional profiles. Without a non-intervention control group, the microbiota differences observed during the 12-week aerobic exercise period can only be considered observational associations, not causal. Additionally, the high dropout rate (72.1%) limits the generalizability of the findings. Clinical trial registration: The Chinese Clinical Trial Registry on 19 October 2022 (ChiCTR2200064801). Full article

Plant extracts and microbiological supernatants were subjected to qualitative and compositional analyses to characterize their bioactive profiles and assess their potential agricultural applications. The garlic (Allium sativum) extract was rich in allicin and selected free amino acids, contained betulin as the dominant triterpene, and displayed a favorable elemental profile with high levels of potassium, phosphorus, sulfur, calcium, and magnesium, with no detectable heavy metals. Detectable amounts of B-group vitamins and vitamin E isoforms were also identified. Qualitative phytochemical screening confirmed the presence of saponins and flavonoids in the garlic extract. The Jerusalem artichoke (Helianthus tuberosus) extract exhibited a significantly higher total phenolic content compared to the garlic extract, with qualitative analysis confirming the presence of saponins, tannins, and flavonoids, suggesting a broader spectrum of bioactive compounds. The two bacterial supernatants were characterized by HPLC analysis and differed in their metabolic profiles: the Enterobacter sp. fermentation broth contained glycerol, 2,3-butanediol, and acetic acid, while the Paenibacillus sp. supernatant additionally contained lactic acid, ethanol, and succinic acid, reflecting distinct fermentation pathways. The in vitro and greenhouse studies aimed to evaluate biological preparations for controlling wheat diseases caused by fungi of the Fusarium genus as well as diseases affecting the stem base. Plant extracts (garlic—Allium sativum, Jerusalem artichoke—Helianthus tuberosus) and supernatants (fermentation broths) obtained with the Paenibacillus and Enterobacter bacteria were tested at three concentrations. In laboratory experiments, the degree of inhibition of the growth of the mycelium of the tested fungal species was determined, while in greenhouse studies, the effectiveness in limiting the development of stem base diseases and the impact of the applied biopreparations on plant growth were evaluated. Among the plant extracts, H. tuberosus demonstrated superior antifungal activity, achieving up to 100% inhibition of R. cerealis mycelial growth at 10% concentration and reducing disease severity by 34.3% compared to the untreated control under greenhouse conditions. Paenibacillus sp. supernatant demonstrated strong in vitro antifungal activity. The results indicate that H. tuberosus extract represents a promising candidate for further field evaluation as a component of sustainable wheat protection programs. Full article

The growing global protein demand and environmental concerns from conventional animal agriculture have driven the exploration of sustainable alternative protein sources. Single-cell proteins (SCPs) from microbial fermentation offer a promising solution. This study comprehensively evaluated the nutritional value and safety profile of SCP produced from Ralstonia eutropha H16 through integrated in vitro and in vivo assessments. Nutritional analyses revealed a high crude protein content of 71.87 ± 5.05 g/100 g dry weight, with total amino acids of 53.67 ± 1.05 g/100 g. The essential amino acid content was 24.38 ± 0.51 g/100 g, accounting for 45% of the total amino acids. An essential amino acid index (EAAI) of 1.46 ± 0.04 and an amino acid score (AAS) of 0.83 ± 0.06 confirmed its classification as a high-quality protein source according to FAO/WHO standards. In vivo rat feeding trials demonstrated an adjusted protein efficiency ratio (PER) of 1.81, exceeding common plant proteins such as wheat (0.8–1.1). True digestibility (TD) reached 85.73%, with a biological value (BV) of 49.37%, net protein utilization (NPU) of 42.33%, and protein digestibility-corrected amino acid score (PDCAAS) of 0.71. Comprehensive safety assessments included chemical contaminant screening, acute oral toxicity studies in rats and mice, in vitro chromosome aberration tests, and erythrocyte micronucleus tests. Heavy metals and aflatoxin B₁ levels were below regulatory limits. Acute oral toxicity studies established LD₅₀ values exceeding 10,000 mg/kg body weight in both rodent species, classifying this protein source as practically non-toxic. The 28-day sub-acute toxicity study showed no significant adverse effects at low doses (6.25% protein replacement). Both genotoxicity assays (mammalian cell chromosome aberration assay and mammalian erythrocyte micronucleus test) returned negative results. These findings establish R. eutropha H16-derived SCP as a safe, nutritious, and sustainable protein source with considerable potential for feed and food applications, contributing to global food security and environmental sustainability. Full article

Tirzepatide, a dual glucagon-like peptide-1 (GLP-1) and glucose-dependent insulinotropic polypeptide (GIP) receptor agonist, demonstrates robust efficacy in glycemic control and weight reduction. However, substantial interindividual variability in treatment response is observed in clinical practice. In this narrative review, we summarize current evidence on clinical, genetic, and molecular predictors of tirzepatide response and discuss their implications for a precision medicine framework. Data from pivotal clinical trials, post hoc analyses, and relevant preclinical and clinical studies were evaluated to identify determinants of glycemic and weight loss responses, as well as hepatic and renal protective effects. Key clinical predictors include tirzepatide dose, duration of diabetes, β-cell function, baseline glycated hemoglobin, sex, age, race, concomitant therapies, and early treatment response. Genetic factors implicated in treatment variability include variants in GLP-1 receptor, GIP receptor, β-arrestin 1, transcription factor 7-like 2, fat mass and obesity-associated protein, and melanocortin 4 receptor, although tirzepatide-specific validation remains limited. Molecular biomarkers such as branched-chain amino acids, insulin-like growth factor–binding protein-1 and -2, the adiponectin-to-leptin ratio, high-sensitivity C-reactive protein, and interleukin-6 show potential as pharmacodynamic indicators of metabolic response. For organ-specific outcomes, procollagen type III N-terminal peptide and magnetic resonance imaging–proton density fat fraction are supported for assessing hepatoprotective effects, while cystatin C–based estimated glomerular filtration rate and urine albumin-to-creatinine ratio are validated markers of renoprotection. Additional candidates—including tumor necrosis factor receptor 1/2, kidney injury molecule-1, and neutrophil gelatinase-associated lipocalin—are promising but require prospective validation. Overall, predicting response to tirzepatide’s multifaceted therapeutic effects necessitates an integrated, multidimensional approach that incorporates clinical characteristics, genetic variation, and molecular profiling. Ongoing validation and harmonization of these predictors may help establish a precision medicine framework for optimizing tirzepatide therapy. Full article

Colombian palm wine is a traditional fermented beverage produced from the sap of Attalea butyracea, whose microbiota and biochemical features remain poorly characterized. A comprehensive analytical framework was applied to palm wine samples from three Andean producers. This included the determination of proximate composition, total phenolic content, and antioxidant activity, alongside a dual microbiological approach: traditional plate counting and high-throughput 16S rRNA/ITS metabarcoding. PICRUSt2 was employed to predict KEGG-based metabolic pathways to elucidate the microbial functional potential. The wines exhibited a low pH (3.35–3.65), a variable ethanol content (1.62–8.40 g/L), high residual sugars, moderate microbial loads, and limited antioxidant activity (as measured using the ABTS and DPPH assays). Analysis using high-throughput sequencing revealed high bacterial community diversity, dominated by Liquorilactobacillus nagelii, Limosilactobacillus fermentum, Limosilactobacillus panis, Lacticaseibacillus casei, and Zymomonas mobilis alongside the yeast Saccharomyces cerevisiae. Functional profiling revealed a significant enrichment in metabolic pathways related to carbohydrates, amino acids, and cofactors/vitamins, as well as xenobiotic biodegradation and metabolism. These findings provide the first integrated microbiological and physicochemical characterization of Colombian palm wine and highlight its biotechnological potential. Full article

The hybrid approach remains a compelling strategy for designing molecules that combine enhanced biological activity with a favorable safety profile. Marine peptides, in particular, have attracted significant attention due to their well-documented broad spectrum of biological activities. Peptides derived from rays have been recognized for their diverse biological activities. Notably, physicochemical properties of these peptides support practical application without requiring further refinement of the mature molecule or specialized formulations. In this study, we present two new chimeric peptides, PK01# and PK02#, which incorporate an opioid pharmacophore linked to a short amino acid sequence derived from the skate Raja porosa. Those compounds interact with the opioidergic system, specifically targeting the mu-opioid receptor (MOR). Furthermore, the compounds were evaluated for their effects on cancer cell viability through in vitro MTT assays (as an exploratory endpoint) and for their binding compatibility with EGFR via in silico docking. Both compounds showed limited effects on cell viability in HeLa, SAS, and PANC-1 cells, while PK02# induced a minor reduction in metabolic activity in glioblastoma cells without reaching IC50 values or significant cytotoxic thresholds. Interestingly, the structures of these hybrid compounds offer valuable insights into the role of phenylalanine residues within their sequences, which appear to be critical for both biological activity and receptor interaction. Moreover, these findings may support future structural optimization of peptide hybrids focused on receptor modulation and biological profiling. Full article

Heart failure with reduced ejection fraction (HFrEF) is increasingly recognized as a systemic metabolic disorder. The aim of this study was to characterize amino acid-related metabolic differences between heart failure with moderately reduced ejection fraction (HFmrEF) (LVEF 40–49%) and HFrEF (LVEF < 40%) and to derive a biologically interpretable composite metabolomic index capable of discriminating between these two stages of systolic dysfunction. We conducted a cross-sectional metabolomic analysis of 42 patients stratified by left ventricular ejection fraction (LVEF < 40% vs. 40–49%). The reference group comprised patients with mildly reduced ejection fraction (LVEF 40–49%), without inclusion of individuals with preserved or normal cardiac function. Targeted amino acid profiling was performed using liquid chromatography-tandem mass spectrometry (LC–MS/MS). Metabolites were standardized and analyzed individually and in combination. A composite index (Heart Failure Amino Acid-Derived Systolic Index: HASI-40), integrating markers of proteolysis and metabolic resilience, was derived to distinguish patients with HFrEF from those with HFmrEF. Discrimination was assessed using receiver operator curve (ROC) analysis with internal validation and multivariable adjustment. Patients with LVEF < 40% exhibited a coordinated metabolic phenotype characterized by reduced methionine, sarcosine, serine, and taurine. While individual metabolites did not retain significance after multiple-testing correction, the composite HASI-40 index remained strongly associated with HFrEF (OR 5.56, 95% CI: 1.70–18.14; p = 0.004), although the wide confidence interval indicates limited precision due to sample size. The index demonstrated good discrimination with an area under the curve (AUC) of 0.862, which improved when combined with age (AUC 0.932). The index represents a standardized composite measure and does not define a diagnostic cutoff for individual patients. These findings suggest that HFmrEF and HFrEF exhibit partially distinct metabolic phenotypes despite overlapping clinical characteristics. These findings suggest that HASI-40 captures metabolic differences between patients with HFmrEF (LVEF 40–49%) and those with HFrEF (LVEF < 40%), reflecting progression toward more advanced systolic dysfunction. However, due to the absence of a control group with preserved ejection fraction, small sample size, and lack of external validation, the index should be considered exploratory and hypothesis-generating rather than clinically applicable. Full article

Over the past decade, an increasing demand for natural antioxidants has driven research into antioxidant peptides and protein hydrolysates from fish and their processing by-products. Catfishes, especially species like Pangasius and Clarias, generate large amounts of protein-rich by-products, which represent a valuable bioresource for valorization. This review discusses advances from the past decade in the production, characterization, and antioxidant capacity of protein hydrolysates and peptides that have been discovered from catfish muscle and by-products. This review emphasizes enzymatic hydrolysis strategies, using Alcalase and other commercial and by-product-derived proteases. Potent antioxidant fractions, particularly those with low molecular weight (<3 kDa) and rich in hydrophobic/aromatic amino acids, have been obtained from the hydrolysates. Mechanisms of antioxidant action, which include hydrogen atom transfer and electron transfer, are discussed in this review, along with the efficacy of catfish-derived antioxidant peptides and protein hydrolysates as demonstrated in chemical and in vivo models. Applications in food systems, such as emulsion-type sausages, have shown potential for shelf-life extension. Nevertheless, knowledge gaps remain, which include an over-dependence on in vitro assays, limited identification of antioxidant peptide sequences, and insufficient data on sensory properties, intestinal permeability, bioavailability, and stability under food processing conditions. Future work should prioritize proteomic characterization, cellular validation, flavor-masking strategies, and scalable production protocols to accelerate the application of catfish protein hydrolysates as viable natural antioxidants for the functional food industry. Full article

Macrophage migration inhibitory factor (MIF) is broadly acknowledged as a central pro-inflammatory regulator, owing to its multifaceted functions including immune cell recruitment, initiation and amplification of pro-inflammatory cytokine cascades, enhancement of macrophage viability, facilitation of macrophage polarization toward a pro-inflammatory state, and attenuation of glucocorticoid-mediated immunosuppression. However, functional investigations of MIF in Mytilus coruscus remain limited. In this study, we identified the MIF gene in M. coruscus, and bioinformatic analyses revealed that the gene encodes a 115-amino-acid polypeptide that exhibits close phylogenetic affinity with MIF homologs from other mollusks. McMIF was predominantly expressed in immune-related tissues, with notably high expression levels in the digestive gland. Upon Vibrio alginolyticus infection, both the mRNA and protein levels of McMIF were significantly upregulated, suggesting that McMIF is involved in the antibacterial immune response of M. coruscus. Meanwhile, the m6A modification level of McMIF was markedly reduced following infection, suggesting a potential relationship between m6A modification and the antibacterial immune function of MIF. Furthermore, knockdown of McMIF followed by LPS stimulation led to an increased level of apoptosis in digestive gland cells, suggesting that McMIF is involved in the inhibition of apoptosis induced by immune stimulation. Collectively, these findings provide insights into the immunological characteristics of McMIF in M. coruscus. Full article

Background/Objectives: Boron neutron capture therapy (BNCT) relies on the selective delivery of boron-10 to tumor cells. Although 4-[¹⁰B]borono-L-phenylalanine (BPA) is currently the only clinically approved BNCT agent, it is limited by poor L-type amino acid transporter 1 (LAT1)/LAT2 selectivity and aqueous solubility. We previously developed 3-borono-5-fluoro-α-methyl-L-phenylalanine (5F-αMe-3BPA), a novel BPA derivative designed to be a LAT1-targeted BNCT/positron emission tomography theranostic agent. This study comprehensively characterizes its pharmacological profile and explores its pharmacokinetic optimization by modulating renal organic anion transporter 1 (OAT1). Methods: Transport kinetics of BPA, related analogs, and 5F-αMe-3BPA were analyzed in HEK293 cells stably expressing LAT1 or LAT2 using Michaelis–Menten analysis. Time-dependent cellular uptake and intracellular retention of BPA and 5F-αMe-3BPA were evaluated in T3M-4 pancreatic cancer cells with or without the LAT1 inhibitor JPH203. In vivo biodistribution was examined in T3M-4 tumor-bearing mice after intravenous administration of 5F-αMe-3BPA or BPA, with assessment of probenecid pretreatment. Results: 5F-αMe-3BPA retained LAT1 affinity comparable to that of BPA while showing markedly reduced LAT2-mediated transport, indicating improved LAT1/LAT2 selectivity. In T3M-4 cells, 5F-αMe-3BPA showed stronger LAT1 dependence, higher steady-state accumulation, and better intracellular retention than BPA under amino acid-containing conditions. Although 5F-αMe-3BPA achieved favorable tumor-to-plasma and tumor-to-muscle ratios in vivo, it was rapidly cleared from circulation. Probenecid pretreatment increased plasma exposure, reduced early renal accumulation, and significantly enhanced tumor boron accumulation, reaching approximately twofold higher levels than control. Conclusions: These findings establish 5F-αMe-3BPA as a highly LAT1-selective BNCT candidate and identify probenecid pretreatment as a clinically translatable pharmacokinetic strategy for maximizing therapeutic boron delivery. Full article

The fruits of Lycium chinense are important medicinal and edible resources with multiple bioactive functions, including hepatoprotective, antioxidant, and immunomodulatory effects. Although this species is widely distributed in China and exhibits abundant germplasm resources, systematic evaluations of fruit quality variation among wild germplasm remain limited, restricting the selection and breeding of superior resources. In this study, eight wild germplasm resources of L. chinense were collected from the eastern coastal regions of China, including Liaocheng (LC), Rugao (RG), Dafeng (DF), Suzhou (SZ), Qidong (QD), Dongtai (DT), Jingjiang (JJ), and Sheyang (SY). A total of 29 fruit quality-related traits, including fruit size, flavonoids, soluble sugars, vitamin C, and amino acids, were analyzed. Significant differences were observed among germplasm resources in both fruit size and internal quality. Comprehensive evaluation based on principal component analysis and entropy weight–grey relational analysis indicated that SY and LC ranked highest. SY exhibited smaller fruits but superior nutritional quality, with higher levels of soluble protein, vitamins, and amino acids. In contrast, LC showed larger fruits and higher contents of polysaccharides and total phenolics, along with stronger antioxidant capacity. Overall, SY and LC represent promising germplasm resources for breeding and utilization of L. chinense. Full article

Medium-resolution cryo-electron microscopy (cryo-EM) density maps preserve substantial information about protein secondary-structure organization; however, accurately recovering the topology and connectivity of α-helices and β-strands remains challenging due to noise, structural heterogeneity, and the intrinsic resolution limitations that obscure residue-level detail. Topology determination is a key intermediate step toward building atomic protein models from medium-resolution cryo-EM density maps. It requires identifying the correct correspondence and orientation between secondary-structure elements (SSEs), i.e., α-helices and β-strands, predicted from the amino-acid sequence and those detected in the three dimensional (3D) density map. Despite significant advances in cryo-EM reconstruction and molecular modelling, this correspondence problem remains a challenging task, particularly in the presence of noisy density maps and in large, topologically complex α/β proteins. To address this issue, we propose a fully automated, classification-based framework that infers protein secondary-structure topology directly from medium-resolution cryo-EM density maps. Specifically, we cast topology determination as a supervised classification problem in three-dimensional space, leveraging geometric learning on model-derived Cα coordinate representations to establish SSE correspondences, and a Dynamic Time Warping (DTW)-based procedure to resolve density-stick directionality. Validation on a benchmark of 38 proteins spanning both simulated and experimental cryo-EM maps and covering diverse fold classes (α, β, and α/β) demonstrates strong and consistent performance. Among the evaluated predictors, the Voronoi (1-NN) classifier achieves the highest average correspondence quality, with a mean F1-score of 96.82% across the full benchmark. The framework also scales to large, topologically dense targets containing up to 65 secondary-structure elements while preserving very fast correspondence inference (<3 ms), offering a substantial improvement over prior baselines in both accuracy and computational cost. Overall, the classification-driven strategy provides reliable SSE-to-density matching and, when coupled with DTW-based direction selection, yields stronger topology constraints that directly support model building and refinement from medium-resolution cryo-EM reconstructions, while remaining easy to integrate into existing structural interpretation pipelines. Full article

Maize (Zea mays L.), a typical thermophilic crop originating from tropical regions, exhibits an inherent sensitivity to low-temperature stress. Cold stress severely restricts maize seed germination, seedling growth, the physiological metabolism, and the final grain yield, which greatly limits its geographical cultivation range and sustainable industrial development. Elucidating the molecular regulatory mechanisms underlying maize cold tolerance and excavating cold-resistant functional genes are essential for the molecular breeding of cold-tolerant maize varieties and expanding maize planting areas in high-latitude and low-temperature-prone regions. In this study, using the strongly cold-tolerant maize inbred line B144 as the experimental material, we cloned the ZmMAPKKKA gene (NCBI accession: LOC103651289) and systematically screened and verified its cold-stress-specific interacting proteins via multiple molecular biological assays. The full-length coding sequence (CDS) of ZmMAPKKKA is 1134 bp, encoding a 377-amino-acid protein with a predicted molecular weight of 40.37 kDa. The quantitative real-time PCR (qRT-PCR) results demonstrated that the ZmMAPKKKA expression was significantly upregulated by 16.56-fold in maize roots after 12 h of low-temperature treatment, indicating a tissue-specific and robust cold response in root tissues. A total of 25 interacting proteins were identified through yeast two-hybrid screening, among which three stress-responsive proteins, including a protein kinase (LOC100286253), a protein phosphatase 2C (PP2C) (LOC542176), and a NAC transcription factor (LOC118474710), were selected for subsequent verification. The Pull-Down, Co-immunoprecipitation (Co-IP), and bimolecular fluorescence complementation (BiFC) assays consistently confirmed that ZmMAPKKKA specifically interacts with these three proteins both in vitro and in vivo under cold stress conditions. This study is the first to construct a ZmMAPKKKA-centered protein interaction module in the maize mitogen-activated protein kinase (MAPK) cascade under cold stress, establishing a novel kinase–phosphatase–transcription factor regulatory cascade that improves the current understanding of cold signal transduction mechanisms in maize. Homologous genes of ZmMAPKKKA in gramineous crops including rice (Oryza sativa) and sorghum (Sorghum bicolor) have been proven to participate in diverse abiotic stress responses, suggesting the conserved functional roles of MAPKKK family genes across gramineous species. Collectively, our findings provide comprehensive insights into the molecular mechanism of the maize MAPK signaling pathway mediating cold stress adaptation and supply valuable functional gene resources for cold-tolerant maize germplasm innovation and molecular breeding. Full article

Nanobodies have emerged as highly valuable biotherapeutic and diagnostic reagents due to their high specificity, low immunogenicity, and superior tissue penetration. However, traditional nanobody discovery methods rely on camelid immunization and phage display techniques, which are time-consuming and labor-intensive. Meanwhile, existing computational prediction methods for Nanobody–Antigen Interaction (NAI) suffer from several limitations: first, general Protein–Protein Interaction (PPI) models cannot adapt to the specific binding patterns of NAI; second, sequence-based models struggle to capture critical binding features, resulting in unsatisfactory prediction accuracy. To address these challenges, we propose an NAI prediction method based on Protein Language Models (PLMs). Specifically, a structure-aware PLM is first fine-tuned on PPI datasets to learn universal protein binding patterns. This model performs joint encoding of amino acid sequences and protein local structure-related sequences. It can implicitly learn spatial structural priors solely from sequence inputs, which alleviates the limitation of conventional sequence-based models in capturing structural binding characteristics. Subsequently, we use mean, max and min pooling to extract complementary global sequence features that a single pooling method cannot fully capture. We then apply voting fusion to reduce prediction bias and improve model robustness under class imbalance and small-sample scenarios. Evaluated on the NAI benchmark dataset constructed from SAbDab-nano, the proposed model outperforms the best baseline methods in key metrics including Accuracy, Recall, F1-score, AUC-ROC, and AUPR. It exhibits robust performance under class imbalance and small-sample scenarios, validating the effectiveness of the framework. Full article

Background: Cancer cells exhibit metabolic reprogramming characterized by increased dependence on glutamine to sustain rapid proliferation and biosynthetic demands. Kidney-type glutaminase (KGA), which catalyzes the first and rate-limiting step of glutamine metabolism, represents a promising therapeutic target, particularly in triple-negative breast cancer (TNBC), an aggressive sub-type lacking effective targeted therapies. This study evaluated 2-amino-4-boronobutyric acid (ABBA), a boronic acid-containing glutamine analog, as a potential KGA inhibitor with anticancer activity. Methods: KGA inhibition was assessed using a fluorometric enzymatic assay. Cytotoxic effects were examined in multiple TNBC cell lines. Covalent docking and molecular simulation analysis were performed to characterize interactions between ABBA and the KGA active site. Results: ABBA potently inhibited KGA activity, with an IC₅₀ of approximately 1.0 μM, demonstrating greater efficacy than several non-proteinogenic amino acid analogs. ABBA induced dose-dependent cytotoxicity across multiple TNBC cell lines, with pronounced sensitivity observed in basal sub-type cells and cellular sensitivity correlated with KGA expression levels. Expression of γ-glutamyl transpeptidase 1 (GGT1) was negligible, and, excluding any off-target effects, the observed anticancer effects are primarily attributed to KGA inhibition. Docking analysis indicated that ABBA forms a reversible covalent adduct with the catalytic Ser286 residue of KGA in a boronate tetrahedral geometry resembling transition-state mimics, while molecular simulation demonstrated stabilization of the complex through hydrogen bonding and electrostatic interactions. Conclusions: ABBA is a potent boron-based glutaminase inhibitor with therapeutic potential for targeting glutamine metabolism in TNBC. Further structural optimization and in vivo evaluation are warranted to advance ABBA toward therapeutic development. Full article